The digene HPV Genotyping PS Test showed high test result agreement in identifying HPV types 16, 18, and 45 in comparison with a validated quantitative PCR HPV Genotyping test. The comparison included 287 PreservCyt and 290 STM archived cervical specimens obtained from a routine screening population. The digene HC2 High-Risk HPV DNA Test result was determined for each specimen prior to inclusion in the study. The results of the digene HC2 High-Risk HPV DNA Test were 238 positive and 52 negative STM specimens and 237 positive and 50 negative PreservCyt specimens. Each specimen was tested for HPV 16, 18, and 45 separately, with both the digene HPV Genotyping PS Test and the qPCR. The total agreement between the digene HPV Genotyping PS Test and the qPCR method was 99.4% (see table).

Agreement between the digene HPV Genotyping PS Test and the qPCR test method

digene HPV Genotyping PS Test

+

–

qPCR +

110

4

qPCR –

7

1610

Total agreement was 99.4%, with a 95% CI of 98.9–99.6%. Each sample was tested in triplicate, with 1720 of 1731 samples showing agreement.

The test agreement of the digene HPV Genotyping PS Test to the qPCR test was determined based on the HPV genotype (see table).

Test result agreement of the digene HPV Genotyping PS Test and the qPCR test for HPV genotype

Genotype

Agreement% (n/N)

95% CI

HPV 16

99.1% (572/577)

98.0–99.6

HPV 18

99.7% (575/577)

98.7–99.9

HPV 45

99.3% (573/577)

98.2–99.7

The test result agreement of the digene HPV Genotyping PS Test to the qPCR test was determined based on the specimen type, with STM specimens showing 99.8% agreement with a 95% CI of 99.2–99.9 and PreservCyt specimens showing 99.0% agreement with a 95% CI of 98.0–99.4.

Using the digene HPV Genotyping PS Test offers analytical specificity with no cross-reactivity between the 3 HPV probe mixes and the following HPV plasmid DNA:

The digene HPV Genotyping PS Test is an in vitro nucleic acid hybridization assay with signal amplification using microplate chemiluminescence for the qualitative detection of the 3 high-risk types of HPV DNA in cervical specimens. The assay is intended as a reflex test to a positive digene HC2 High-Risk HPV DNA Test result, to determine the presence or absence of 16, 18, or 45 genotypes.

The digene HPV Genotyping PS Test, performed in triplicate for each cervical specimen, genotypes for 3 high-risk types (16, 18, and 45) of HPV DNA in cervical specimens. After denaturation, specimens are hybridized with specific HPV RNA probes which target the full HPV genome. The resultant RNA:DNA hybrids are captured onto the surface of a microplate well coated with antibodies specific for RNA:DNA hybrids. In the digene HPV Genotyping PS Test, the formation of hybrids and their subsequent capture onto the surface of the microplate occur simultaneously during a single step. Immobilized hybrids then react with specific alkaline phosphotase-conjugated and are detected with a chemiluminescent substrate using the DML 2000 or DML 3000 instrument.

Applications

The digene HPV Genotyping PS Test is a reflex test intended for the qualitative detection of high-risk HPV types 16, 18, and 45 following a positive digene HC2 High-Risk HPV DNA Test result. The digene HPV Genotyping PS Test may be used for testing women in cervical cancer screening programs.